Diagnosis of Bovine Anaplasmosis Using Anaplasma Antigenic Markers Identified by Western Blotting
Kwak, Young Ro
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https://hdl.handle.net/2142/71354
Description
Title
Diagnosis of Bovine Anaplasmosis Using Anaplasma Antigenic Markers Identified by Western Blotting
Author(s)
Kwak, Young Ro
Issue Date
1988
Doctoral Committee Chair(s)
Smith, Ronald D.
Department of Study
Veterinary Medical Science
Discipline
Veterinary Medical Science
Degree Granting Institution
University of Illinois at Urbana-Champaign
Degree Name
Ph.D.
Degree Level
Dissertation
Keyword(s)
Biology, Veterinary Science
Abstract
Diagnosis of bovine anaplasmosis relies on serology using crude parasite extracts, detection of parasitemia, or subinoculation of blood from suspected animals into susceptible animals. Anaplasma can not be cultured in vitro. The feasibility of using antigenic markers detected by Western blotting (WB) for diagnosis of bovine anaplasmosis was studied.
Antigenic markers of diagnostic and taxonomic value were identified by using an antigen bank of 5 different Anaplasma organisms and 6 different antisera to 6 distinct Anaplasma. These markers were conserved after passage of bovine Anaplasma through white-tailed deer. This implies that white-tailed deer may serve as reservoirs for bovine anaplasmosis without signficant alteration of antigenic markers, and that WB is useful to identify the markers regardless of the source of bovine infection. Recognition of antigenic markers was stable with little animal variation regardless of age of the animal and severity of the disease in both cross sectional and longitudinal studies. The results from experimentally-infected animals were used to determine the stage of infection (incubation, sick, convalescent, and carrier stages) of field cases with hematologic or serologic evidence of naturally-acquired bovine anaplasmosis. Concordance between WB and rapid card agglutination test (RCAT) was 99% in cattle from a ranch where a field outbreak occurred. When results from WB were compared with those from complement fixation test (CFT), CFT titers did not correspond to the intensity of antigen bands on blots. Concordance was 93% between WB and RCAT, and 60% between WB and indirect fluorescent antibody test (IFAT) in naturally-infected carrier animals from a second ranch.
In conclusion, Western blotting was able to identify Anaplasma antigenic markers regardless of the source of bovine infection, with little variation regardless of age of animals and severity of the disease. This technique was useful for diagnosis and clinical staging of anaplasmosis cases, and was validated as a diagnostic test by comparing results with CFT, IFAT and RCAT.
Further research to develop a synthetic or cloned peptide immunoassay using specific markers may lead to more accurate serologic tests for rapid diagnosis of the disease.
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